Human Reproduction, Vol. 16, No. 6, 1200-1203,
June 2001
© 2001 European Society of Human Reproduction and Embryology
Evidence of reduced single-stranded testicular sperm DNA from obstructive azoospermic men after 3 days of in-vitro culture
1 Fertility Clinic and 2 Laboratory of Biology and Psychology of Human Fertility, Erasmus Hospital French Speaking Free University of Brussels, 808, Route de Lennik, B1070 Brussels, Belgium
The aim of the present study was to verify whether culturing testicular tissue, to obtain a higher percentage of motile spermatozoa and a better post-thaw recovery rate, affected the ratio between single/double-stranded sperm DNA and, consequently, DNA sensitivity to damage. Testicular biopsy samples from men with obstructive and secretory azoospermia, candidates for assisted reproductive treatment, were cultured for 72 h. The percentage of motile spermatozoa and the single/double stranded DNA ratio were assessed on the day of retrieval (day 0) and again on day 3. The single/double stranded DNA ratio was measured by the acridine orange (AO) staining method. Spermatozoa were classified as green (double-stranded chromatin) or red fluorescing (single-stranded chromatin). In obstructive azoospermia, median motility was 22% (range 10–44%) on day 0 and 50% (range 38–63%) on day 3 (P < 0.01). The median percentage of red stained spermatozoa was 53.5% (range 0.1–88%) on day 0 and 20% (range 2.7–99.9%) on day 3 (P < 0.05). No changes were observed in secretory azoospermia. The culture procedure from obstructive azoospermia not only increased the post-thaw recovery rate, as previously observed, but also reduced the portion of spermatozoa containing single-stranded DNA, thereby increasing the availability of double-stranded DNA spermatozoa for ICSI use.
Key words: acridine orange/azoospermia/in-vitro culture/sperm DNA/testicular sperm extraction
3 To whom correspondence should be addressed at: Fertility Clinic, Department of Obstetrics and Gynaecology, Erasmus Hospital, Route de Lennik 808, 1070 Brussels, Belgium. E-mail: semilian{at}ulb.ac.be
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